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Goodwin et al, Biophys J. 2005, 89(2): 1398 Diffusional mobilities of GFP-HRas, GFP-NRas, and GFP-KRas are similar to one another and the fluorescent lipid probes DiIC16 and DiIC18 in the plasma membrane of COS-7 cells Fluorescencce Recovery After Photobleaching

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~ 400 nm Patterson at al. Science 2002, 297: 1873 Photoconversion in wild-type GFP is thought to involve a shift in the chromophore population from the neutral phenolic form to the anionic phenolate form Photoconversion Molecular Basis

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Keese et al. J Biol Chem 2005; 280: Confocal imaging of EGFR phosphorylation in a SW-480 cell by acceptor photobleaching FRET. a, F4-Cy3 staining before photobleaching. b, Py72-Cy5 staining before photobleaching. c, F4-Cy3 staining after photobleaching of Cy5. The area in which Cy5 was photobleached is marked. Here an increase in the intensity of the Cy3 fluorescence can be observed. d, Py72-Cy5 staining after photobleaching. The area in which Cy5 was photobleached is marked. e, difference image of c and a (I(donor postbleach) - I(donor prebleach)) showing the increase in the Cy3 fluorescence. Acceptor Photobleaching FRET

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Chromophore-assisted laser inactivation Causing local damage to a protein of interest Rajfur Z. et al.Nat Cell Biol. 2002:4; 286 Figure3 A typical example of ‘slow’ retraction of stress fibre after CALI irradiation at the FA. a–f,The stress fibres are visualized as bright green spots of expressed EGFP– α -actinin. The irradiated spot is indicated by the yellow circle. The yellow arrowhead represents a reliablemark for the initial position of the FA and the red arrowhead follows the position of retracted stress fibre after CALI treatment on the subsequent panels.